[A new automated haematology analyser: the Excell 2280]. / Un nouvel automate de cytologie hématologique: l'Excell 2280.

Excell 2280 analyser is a new automated haematology analyser manufactured by Drew Scientific Inc, Texas, USA, and distributed in France by MAXMAT S.A., Montpellier. It can achieve 80 complete blood cell counts per hour, with leukocyte differential counts. Three sampling possibilities are included: a direct one (open tubes, 180 microL), a blood saver one (80 microL) and an automatic, through-the-cap one (180 microL). The analytic principles are: electrical impedance for cell counting (WBC, RBC, platelets, MCV) and RBC/platelet sizing; and a new multidimensional optical system using a laser light scattering flow cytometer for WBC counting and classification. We evaluated the Excell 2280 in our laboratory: we quantified intra-run and within-run variations, correlations between the automatic and the direct sampling method, stability of the results over time, linearity of the detections and finally correlation between results obtained with this analyzer and the Gen'S one from Beckman-Coulter Inc. The obtained results were within the theoretical ranges given by the manufacturer. The presence of any abnormal result, or of any flag, must systematically lead to check the blood smear. This new automated haematology analyser appears to be convenient for emergency room-related laboratories, and for routine small-to-medium laboratories.

Reliability of hemostasis biomarkers is affected by time-dependent intra-patient variability.

Essentials Nucleosomes and free DNA are two newly described biomarkers in venous thromboembolism (VTE). Reliability of nucleosomes, plasma free DNA and conventional hemostasis markers were studied. Hemostasis biological parameters vary over a short time-frame in VTE patients. Hemostasis biological parameters also vary over a short time-frame in healthy controls. SUMMARY: Background Previous studies have associated neutrophil-derived circulating nucleosomes and plasma free DNA with venous thromboembolism (VTE). However, there are few data concerning these two biomarkers and no studies have compared the reliability of nucleosomes and plasma free DNA against that of conventional hemostasis markers. Objectives We performed a 3-year prospective study of nucleosomes and plasma free DNA levels in comparison with conventional hemostatic biomarkers and blood cells. Patients/Methods Fifteen healthy controls and 22 randomly selected patients with a history of VTE were followed monthly for 6 months. The reliability of these markers was evaluated by the intraclass correlation coefficient (ICCs). Results and Conclusions In healthy controls and patients, we found a low reliability for nucleosomes and plasma free DNA, with ICCs at 0.538 (95% confidence interval [CI], 0.334-0.764) and 0.091 (95% CI, -0.026-0.328), respectively, in the healthy controls, and at 0.213 (95% CI, 0.042-0.463) and 0.161 (CI 95%, 0.008-0.398) in the patient group. For the conventional hemostasis biomarkers and for blood cells, reliability ranged from poor to good in the healthy volunteers and from poor to acceptable in the patient group. Our study shows for the first time that hemostasis biological parameters spontaneously vary over a short time-frame in VTE patients and, more surprisingly, in normal individuals. The clinical value of such intra-individual variations is currently unknown. This variability might mean reinterpreting diagnostic or prognostic models based on static evaluation of individuals. Studying the intrinsic value of individual patterns of markers' variability is warranted.

Systematic mutation of bacteriophage T4 lysozyme.

Amber mutations were introduced into every codon (except the initiating AUG) of the bacteriophage T4 lysozyme gene. The amber alleles were introduced into a bacteriophage P22 hybrid, called P22 e416, in which the normal P22 lysozyme gene is replaced by its T4 homologue, and which consequently depends upon T4 lysozyme for its ability to form a plaque. The resulting amber mutants were tested for plaque formation on amber suppressor strains of Salmonella typhimurium. Experiments with other hybrid phages engineered to produce different amounts of wild-type T4 lysozyme have shown that, to score as deleterious, a mutation must reduce lysozyme activity to less than 3% of that produced by wild-type P22 e416. Plating the collection of amber mutants covering 163 of the 164 codons of T4 lysozyme, on 13 suppressor strains that each insert a different amino acid substitutions at every position in the protein (except the first). Of the resulting 2015 single amino acid substitutions in T4 lysozyme, 328 were found to be sufficiently deleterious to inhibit plaque formation. More than half (55%) of the positions in the protein tolerated all substitutions examined. Among (N-terminal) amber fragments, only those of 161 or more residues are active. The effects of many of the deleterious substitutions are interpretable in light of the known structure of T4 lysozyme. Residues in the molecule that are refractory to replacements generally have solvent-inaccessible side-chains; the catalytic Glu11 and Asp20 residues are notable exceptions. Especially sensitive sites include residues involved in buried salt bridges near the catalytic site (Asp10, Arg145 and Arg148) and a few others that may have critical structural roles (Gly30, Trp138 and Tyr161).

Alteration of T4 lysozyme structure by second-site reversion of deleterious mutations.

Mutations that suppress the defects introduced into T4 lysozyme by single amino acid substitutions were isolated and characterized. Among 53 primary sites surveyed, 8 yielded second-site revertants; a total of 18 different mutants were obtained. Most of the restorative mutations exerted global effects, generally increasing lysozyme function in a number of primary mutant contexts. Six of them were more specific, suppressing only certain specific deleterious primary substitutions, or diminishing the function of lysozymes bearing otherwise nondeleterious primary substitutions. Some variants of proteins bearing primary substitutions at the positions of Asp 20 and Ala 98 are inferred to have significantly altered structures.

Polypeptide composition of an adenovirus type 5 used in cancer gene therapy.

For cancer gene therapy, a recombinant adenovirus serotype 5 named RPR/INGN201 has been constructed by susbtitution of the E1 region with human tumor suppressor gene p53. The protein components of RPR/INGN201 virions were separated by reversed-phase HPLC and were individually identified by electrospray time-of-flight mass spectrometry and N-terminal sequencing, both on intact proteins and on their proteolytic fragments after trypsin digestion. Twenty-five peptide components of the proteome (including fiber) with greater than 0.25-0.5% contribution to the protein content of the virus were identified and characterized. Fiber was confirmed to be partially glycosylated (both the non-glycosylated and the monoglycosylated states were identified), and two proteins were isolated and identified as phosphorylation derivatives, namely protein V (non-phosphorylated and monophosphorylated) and protein IIIa (mono- and diphosphorylated). This new analytical tool proved to be very useful not only for refining our current knowledge of the polypeptide repertoire of purified infectious virions but also for monitoring and very rapidly identifying structural modifications resulting from changes in the manufacturing process. It was also used successfully for the characterization of various adenoviral constructs.

Role of laparoscopy in blunt perforations of the small bowel.

BACKGROUND: The purpose of this study was to determine whether laparoscopy could improve our ability to diagnose and treat perforations of the small bowel. METHODS: From 1985 to 2001, among 250 patients admitted for a blunt abdominal trauma, 195 surgical explorations were performed, comprising 42 laparoscopies. On admission, 108 patients underwent an abdominal ultrasonography (US) and 104 a computed tomography (CT). Thirty-nine patients had a blunt small bowel trauma (BSBT) that was explored and/or treated by laparoscopy (n = 15) or celiotomy (n = 24). We compared the patients who underwent emergency surgery (23) and those who were operated on after a mean delay of 51 h. RESULTS: On admission, for detecting free intraperitoneal fluid with a BSBT, the sensitivity of CT was 93.3% and specificity was 13.6%. Regarding the 23 patients who underwent emergency surgery, 8/20 had positive US and 10/15 had positive CT, whereas the 15 celiotomies and 8 laparoscopies indicated BSBT. Regarding the 16 patients who underwent delayed surgery, 9 patients underwent a celiotomy and 7 a laparoscopy; all BSBTs were diagnosed. In searching for BSBT, the sensitivity and specificity of laparoscopy were both 100%, whereas the sensitivity of CT was 83.3% and specificity was 22.2%. Twenty of 23 patients undergoing emergency surgery had bowel suture repair compared to 3/16 after delayed surgery. In contrast, a resection was necessary in 10/12 patients treated after a delay compared to 2/17 operated in emergency. Five BSBTs were sutured by laparoscopy, whereas 10 required conversion. Laparoscopy avoided celiotomy in 15 cases. There were no deaths and no enterostomy. CONCLUSION: In hemodynamically stable patients with blunt abdominal trauma, laparoscopy safely and effectively identifies small bowel injuries. Early recognition of these injuries and timely surgical treatment offer the best prognosis.

[Well-differentiated papillary mesothelioma of the peritoneum: an attenuated malignant tumor. Review of the literature apropos of a case]. / Le mésothéliome péritonéal papillaire bien différencié: une tumeur à malignité atténuée. Revue de la littérature à propos d'un cas.

The authors present a new case of a well-differentiated papillary mesothelioma of the peritoneum. This is an uncommon tumor which have a slow evolution like a low malignant potential tumor. But, because of its tendency to recurrence, the designation of Well-Differentiated Tumor is better. The diagnosis with others peritoneal tumors is sometimes difficult, especially with the Peritoneal Serous Tumors. Tumor recurrence must be treated by curative surgery. Adjuvant therapy is discussed for the diffuse form.

[Causes and risk factors in post-neonatal mortality in France in 1987 according to a national survey]. / Causes et facteurs de risque de la mortalité postnéonatale en France en 1987, d'après une enquête nationale.

A stagnation in postneonatal mortality (28 days-1 year) is reported in France as in other similar countries. More than fifty per cent of these deaths are registered under the sudden infant death syndrome, a cause which is classified in the symptoms and ill-defined conditions chapter of the International Classification of Diseases. To establish the actual rate of sudden infant death syndrome (SIDS) it was necessary to carry out a specific retrospective survey. All postneonatal deaths (3917) registered in France from September 1986 to December 1987 were studied by means of questionnaires sent to the certifying doctor and to the families. The sudden infant death rate is estimated at 1.2 per 1,000 and the risk factors of the total postneonatal deaths are birthweight, prematurity, transfer into neonatal care unit, and age of the mother.

[Evaluation of an automated method for measuring low-density lipoprotein cholesterol]. / Evaluation du dosage direct du cholestérol des lipoprotéines LDL.

We evaluated an automated method for measuring low-density lipoprotein cholesterol (LDL-cholesterol) with Integra 800 (Roche Diagnostics). This method uses detergents which show selectivity for the different lipoproteins. Our evaluation has shown a within run imprecision of 2% and a between-assay imprecision of 1.2%. This method was not perturbed by haemoglobin, bilirubin or lipids. However, the reagents remain expensive: we will use it when calculation or interpretation of lipid results are not possible.

[Prosthetic treatment of bilateral inguinal hernias via laparoscopic approach or Stoppa procedure]. / Traitement prothétique des hernies inguinales bilatérales par voie laparoscopique ou par opération de Stoppa.

AIM OF THE STUDY: The aim of this prospective non-randomized study was to compare Stoppa's technique to laparoscopic approach in totally extraperitoneal repair of bilateral inguinal hernia. PATIENTS AND METHOD: From December 1996 to December 1998, 117 consecutive patients with 234 hernias underwent either Stoppa's technique (74 patients) or a totally extraperitoneal laparoscopic approach (43 patients). Patients were randomized in two groups according to the surgeon to whom they were referred. All patients were reviewed in December 1999. RESULTS: There was no mortality. Complications occurred in 3% of patients after Stoppa's technique (group S) and in 4% of patients in the laparoscopic group (group L). The conversion rate was 7% (3 cases). Postoperative analgesia use, hospital stay, and duration of disability were significantly shorter in group L, the cost was lower, but the operating time was significantly longer than in group S. Recurrence rates were similar in the two groups: 2% in group S, 1.1% in group L. CONCLUSION: The laparoscopic approach appears to be preferable to Stoppa's technique in the treatment of bilateral inguinal hernia.

[Excision of the duodenojejunal angle for tumor. From simple excision to locoregional excision]. / Résection de l'angle duodéno-jéjunal pour tumeur. De l'exérèse simple à l'exérèse loco-régionale.

Locoregional extensions are often observed in tumors of the duodenojejunal angle, either because of their anatomic situation or in cases diagnosed late. Endoscopic and radiographic explorations (basically abdominal CT scan) are indicated to search determine the degree of extensions to neighboring organs and vessels. Colic and mesocolic involvement are at the limit of preoperative explorations. Based on 4 observations of increasing complexity, we describe our operative strategy allowing tumor exeresis in a single block. Extension to the superior mesenteric artery is the main contraindication for exeresis.

[Cystic lymphangiomas of the colon. Apropos of 2 cases]. / Les lymphangiomes kystiques du côlon. A propos de 2 observations.

The authors report about two observations of cystic lymphangioma of the right colon. The patients complained of intermittent abdominal pain. Preoperative diagnosis is based on endoscopic findings (2 cases) and computed tomography films (1 case). A colonic carcinoma is associated in one case. Endoscopic polypectomy will be curative for pedunculated or small lymphangiomas. In other cases segmental resection seems to be the treatment of choice.

[Cancer of the penis: the value of systematic biopsy of the superficial inguinal lymph nodes in clinical N0 stage patients]. / Cancer du pénis: valeur de la biopsie systématique des ganglions inguinaux superficiels chez les patients N0 clinique.

From 1985 to 1991, bilateral surgical biopsy of the superomedial group of superficial inguinal lymph nodes, considered to be the first draining nodes, was performed in 24 patients with clinical stage T1-3 N0 M0 squamous cell carcinoma of the penis at the same time as surgical treatment of the primary. This procedure was technically successful in every case, but no lymph node metastases were detected. As bilateral biopsy was negative in these 24 patients, clinical surveillance was implemented: review every 2 months for 2 years and self-palpation by the patient. Seven patients (29.1%) developed one or more suspicious ilio-inguinal lymph nodes after a mean interval of 11.85 +/- 8.02 months: 1) In 6 patients, bilateral ilio-inguinal lymph node dissection was then performed, confirming the neoplastic nature of the inguinal node metastases without iliac metastases in 3 of these patients. 2) As the remaining patient presented with unilateral iliac node metastases, proven histologically by surgical biopsy, systemic chemotherapy was introduced prior to ipsilateral inguinal lymph node dissection. This finding questions the theory according to which the sentinel superomedial inguinal node constitutes the first draining node and demonstrates the existence of several pathways of lymphatic drainage towards superficial and deep inguinal nodes. This biopsy, which was always negative in our series, is insufficient to guide our therapeutic approach in clinical N0 patients. The decreased complication rate following inguinal lymph node dissection should certainly encourage us to prefer surgery, particularly superficial inguinal lymph node dissection with preservation of the great saphenous vein.

Reflection on the measurement and use of the topography of the indentation imprint.

The goal of this paper is to study the main uses of the residual imprint of the indentation test. It also discusses the different technologies and methods employed in this context. The difficulties encountered when trying to exploit the full potentials of the imprint are thoroughly examined. A survey of the literature on the quantification of the pile-up clearly shows that there is a lack of consensus on the measurement of the residual imprint as well as on treatment methods. Therefore, in order to widen the application fields of the indentation residual imprint, relevant and standardized indicators should be established.

Second-site reversion of a structural defect in bacteriophage T4 lysozyme.

A plasmid-borne gene encoding bacteriophage T4 lysozyme with a structural mutation, Tyr161-Ala, was mutagenized by by the use of polymerase chain reaction. The mutagenized gene was inserted into a specialized bacteriophage lambda cloning vector that must acquire a functional lysozyme gene in order to form plaques. Functional variants of the mutant lysozyme were selected. Three compensatory second-site revertants were obtained: Thr152-Met, Lys43-Ile, and Thr151-Ala. The effects of these mutations are interpreted in light of previous structural and genetic studies of T4 lysozyme.

Epidemiology of schistosomiasis in the riverine population of Lagdo Lake, northern Cameroon: mixed infections and ethnic factors.

The distribution of S. mansoni and S. haematobium was evaluated using a cluster sampling technique in 15 fishing villages around Lagdo lake and 2 inland villages in northern Cameroon. The overall prevalence of S. haematobium infection was 26% and of S. mansoni was 15%, while 7% of the population sampled had mixed infections. The highest prevalences were observed in larger villages (greater than 200 persons) where sanitary conditions were poor. The intensity of S. haematobium infection in those also infected with S. mansoni was two to three times greater than in those with S. haematobium alone. Differences in prevalence among ethnic groups was attributed to occupational and cultural habits. The factors leading to the diversity of the distribution of schistosomiasis within a man-made lake are discussed.

Functional significance of conserved amino acid residues.

A systemic study of single amino acid substitutions in bacteriophage T4 lysozyme permitted a test of the concept that conserved amino acid residues are more functionally important than nonconserved residues. Substitutions of amino acid residues that are conserved among five bacteriophage-encoded lysozymes were found to lead more frequently to loss of function than substitutions of nonconserved residues. Of 163 residues tested, only 74 (45%) are sensitive to at least one substitution; however, all 14 residues that are fully conserved are sensitive to substitutions.

Constitutive mutants in the yeast pheromone response: ordered function of the gene products.

The alpha factor pheromone inhibits the division of yeast a cells. A general method was developed for isolating mutants that exhibit constitutive activation of the pheromone response pathway. A dominant allele of the STE4 locus was recovered in addition to recessive mutations in the SCG1 gene. SCG1 and STE4 are known to encode G alpha and G beta homologs, respectively. Analysis of double mutants suggests that the STE4 gene product functions after the SCG1 product but before the STE5 product.

Epidemiology, anthropology and health education.

Epidemiology gives only a partial picture of a health problem. Without a knowledge of social, cultural, and economic conditions as well, one can fall into farcical errors in designing interventions.

PIP: A large dam was built on the Benoue River in the north of Cameroon in 1982, creating a reservoir of over 500 km. The health care system could no longer meet day-to-day needs. The principles of schistosomiasis control are simple. However, implementing them raises a lot of problems. In 1986, an epidemiological survey was carried out, covering 15 villages of about 1000 people chosen at random. At the same time, a socio anthropological study was done in certain representative villages. Interviews were conducted to gather information on perceptions of health, disease, socio cultural structures, and the extent to which the natives understood the educational imagery that was used. Excreters of schistosoma eggs were detected. Overall prevalence rates were 25% for urinary schistosomiasis, and 15% for Mason's intestinal schistosomiasis, with ranges of 7-35% and 7-29%, respectively. 70% of the egg excreters were under 20 years old. 3 high prevalence villages were large in size and had primitive sanitation. Patients can be treated with praziquantel, which is effective against Schistosoma, hematobium and Schistosoma mansoni. Good environmental surveillance and a good sanitation system are necessary. From the anthropological survey, information was gathered so that a community-centered system for monitoring and preventing schistosomiasis might be established. A number of different tribes are represented, and the nature of settlements is transitory. The villagers usually said that schistosomiasis came from the drinking of lake water or sexual intercourse. Most groups urinate or defecate away from built-up areas. Illness was usually linked with pain. 2 types of treatment are available for schistosomiasis. The 1st is traditional, the 2nd involves attending a health center. The schistosomiasis parasite cycle was not well understood by many villagers, revealing the need for simple educational messages.